Purification and primary sequences of the major arginine-containing antifreeze glycopeptides from the fish Eleginus gracilis

• Published in: The Journal of biological chemistry Vol. 261; no. 14; pp. 6384 - 6389
• Main Authors: Burcham, T S, Osuga, D T, Rao, B N, Bush, C A, Feeney, R E
• Format: Journal Article
• Language: English
• Published: Bethesda, MD Elsevier Inc 15.05.1986; American Society for Biochemistry and Molecular Biology
• Subjects: Amino Acid Sequence; Aminoacids, peptides. Hormones. Neuropeptides; Analytical, structural and metabolic biochemistry; Animals; Antifreeze Proteins; Arginine - analysis; Biological and medical sciences; Fishes; Fundamental and applied biological sciences. Psychology; Glycoproteins - analysis; Glycoproteins - isolation & purification; Proteins; Vertebrata; Purification; Glycopeptide; Animal; Pisces; Primary structure; Antifreeze protein
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/S0021-9258(19)84573-8

An Arg-containing antifreeze glycoprotein from the polar fish Eleginus gracilis was isolated, and the major components were purified to homogeneity. The general protocol for purification was chromatography of serum on DEAE-cellulose, followed by chromatography on a cation exchanger. DEAE-cellulose chromatography resulted in two fractions, A and B. Fraction A contained most of the antifreeze glycoprotein found in E. gracilis (approximately 80% by weight) and consisted of 13 distinct components. Unlike antifreeze glycoproteins from other previously studied polar fish, Fraction A contained both low and high molecular weight antifreeze glycoprotein components. The two major components of Fraction A were sequenced and compared with the sequence of antifreeze glycoproteins 7 and 8 from both Boreogadus saida and Pagothenia borchgrevinki. The antifreeze glycoproteins from E. gracilis were shown to have a similar composition to those previously studied, except for an additional Ala-Arg dipeptide at the carbon terminal in the major components of Fraction A and the position of Pro in the low molecular weight components. The activity of E. gracilis antifreeze glycoproteins is the subject of a companion article (Burcham, T. S., Osuga, D. T., Yeh, Y., and Feeney, R. E. (1986) J. Biol. Chem. 261, 6390-6397).